Figure 3
Figure 3. Correlative fluorescence and scanning electron microscopy of VWF dots. HUVECs were stimulated for 20 minutes with 80nM PMA in the presence of FITC-conjugated anti-VWF antibodies to detect extracellularly accessible VWF. (A-A′) Overlay of a correlated VWF fluorescence and scanning electron microscopy (SEM) micrograph of part of a HUVEC, revealing that the VWF immunofluorescence (in green) corresponds to globular structures on the cell surface. Scale bar represents 5 μm. (A′) SEM micrograph of the same area shown in panel A. Cell surface areas containing clusters of VWF dots often contained one or more pores (open arrowhead). (B) Higher magnification of a cluster of VWF dots indicated by the black arrowhead in panel A′. Scale bar represents 500 nm. (C) Higher magnification of the pore indicated by the open arrowhead in panel A′. Notice the clearly defined rim of the pore. Scale bar represents 500 nm. (D-D') When anti-VWF antibodies are omitted during secretagogue challenge, long VWF strings are found on the surface of the cells. (D) Typical example of branching VWF strings (black arrowheads) forming a network. Note the pore (white arrowhead) from which a VWF string emanates. Scale bar represents 1 μm. (D′) Close-up of the secretion pore shown in panel D. Scale bar represents 500 nm. (E) Close-up of another secretion pore releasing globular material and a VWF string (white arrowhead). Pore in the membrane showing globular material and a filament (white arrowhead). The filament appears to arise from the globule. Small filamentous material is also present in the pore (arrow). Scale bar represents 500 nm. (F) High magnification view of a VWF string and associated globule. The string (arrowhead) consists of several entwined filaments (arrows) forming a bundle. Scale bar represents 500 nm. The fluorescence micrograph was acquired using a Leica SP2 confocal laser-scanning microscope and a 63× oil-immersion objective with a numeric aperture of 1.33 (Leica PL APO). Scanning electron micrographs were made with a JEOL JSM-6700F field emission scanning electron microscope at 5.0 kV.

Correlative fluorescence and scanning electron microscopy of VWF dots. HUVECs were stimulated for 20 minutes with 80nM PMA in the presence of FITC-conjugated anti-VWF antibodies to detect extracellularly accessible VWF. (A-A′) Overlay of a correlated VWF fluorescence and scanning electron microscopy (SEM) micrograph of part of a HUVEC, revealing that the VWF immunofluorescence (in green) corresponds to globular structures on the cell surface. Scale bar represents 5 μm. (A′) SEM micrograph of the same area shown in panel A. Cell surface areas containing clusters of VWF dots often contained one or more pores (open arrowhead). (B) Higher magnification of a cluster of VWF dots indicated by the black arrowhead in panel A′. Scale bar represents 500 nm. (C) Higher magnification of the pore indicated by the open arrowhead in panel A′. Notice the clearly defined rim of the pore. Scale bar represents 500 nm. (D-D') When anti-VWF antibodies are omitted during secretagogue challenge, long VWF strings are found on the surface of the cells. (D) Typical example of branching VWF strings (black arrowheads) forming a network. Note the pore (white arrowhead) from which a VWF string emanates. Scale bar represents 1 μm. (D′) Close-up of the secretion pore shown in panel D. Scale bar represents 500 nm. (E) Close-up of another secretion pore releasing globular material and a VWF string (white arrowhead). Pore in the membrane showing globular material and a filament (white arrowhead). The filament appears to arise from the globule. Small filamentous material is also present in the pore (arrow). Scale bar represents 500 nm. (F) High magnification view of a VWF string and associated globule. The string (arrowhead) consists of several entwined filaments (arrows) forming a bundle. Scale bar represents 500 nm. The fluorescence micrograph was acquired using a Leica SP2 confocal laser-scanning microscope and a 63× oil-immersion objective with a numeric aperture of 1.33 (Leica PL APO). Scanning electron micrographs were made with a JEOL JSM-6700F field emission scanning electron microscope at 5.0 kV.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal